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首页> 外文期刊>Allergy >Identification of a serine protease as a major allergen (Per a 10) of Periplaneta americana.
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Identification of a serine protease as a major allergen (Per a 10) of Periplaneta americana.

机译:将丝氨酸蛋白酶鉴定为美洲大plane的主要变应原(每10个)。

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BACKGROUND: Cockroach allergens are associated with the development of asthma, but none of these has been characterized for proteolytic activity. This study was undertaken to isolate and characterize a protease from Periplaneta americana and determine its allergenicity. METHODS: A serine protease was isolated from P. americana extract using benzamidine sepharose column and characterized by immunobiochemical methods. Allergenicity of the protease was assessed by enzyme-linked immunosorbent assay, immunoblot, intradermal testing, histamine release and peripheral blood mononuclear cells (PBMCs) proliferation. RESULTS: Affinity purified protein of approximately 28 kDa (Per a 10) showed a single band of activity in gelatin zymogram and agarose plate assay. N-terminal sequence (IVGGRPAQI) revealed similarity with mite serine protease allergens and insect trypsins. It demonstrated proteolytic activity with azocollagen > gelatin > defatted-milk > casein including serine protease specific substrate, N-benzoyl-arginine-ethyl-ester-hydrochloride. It was inhibited by serine protease inhibitors, namely aprotinin > pefabloc > AEBSF > PMSF > benzamidine > antipain > leupeptin and trypsin-specific inhibitor (tosyl-lysyl-chloromethyl-ketone) suggesting it to be a trypsin-like serine protease. Per a 10 was recognized as a major allergen, showing IgE reactivity with >80% of cockroach sensitized patients by skin tests and immunoblot. It could induce significant histamine release (P < 0.05) in blood and secretion of interleukin-4 (IL-4) (P < 0.05) and IL-5 (P < 0.05) in culture supernatant of PBMCs from cockroach hypersensitive patients, suggesting a strong allergenic potency. CONCLUSION: A serine protease isolated from P. americana was demonstrated to be a major allergen (Per a 10). It has a potential for component-based diagnosis of allergy and will be useful in elucidating the mechanism of allergy.
机译:背景:蟑螂过敏原与哮喘的发展有关,但这些蛋白原均没有蛋白水解活性。进行这项研究以分离和表征美洲原i的蛋白酶并确定其变应原性。方法:采用苯甲sep琼脂糖凝胶柱从美洲豹提取物中分离出丝氨酸蛋白酶,并进行免疫生化分析。通过酶联免疫吸附测定,免疫印迹,皮内测试,组胺释放和外周血单核细胞(PBMC)增殖来评估蛋白酶的变应原性。结果:亲和纯化的大约28 kDa的蛋白质(每10个)在明胶酶谱图和琼脂糖平板测定中显示出一条单一的活性带。 N末端序列(IVGGRPAQI)显示与螨丝氨酸蛋白酶过敏原和昆虫胰蛋白酶相似。它表现出的蛋白水解活性包括偶氮胶原>明胶>脱脂牛奶>酪蛋白,其中包括丝氨酸蛋白酶特异性底物,N-苯甲酰基-精氨酸-乙基酯-盐酸盐。它被丝氨酸蛋白酶抑制剂抑制,即抑肽酶> pefabloc> AEBSF> PMSF>苄am>抗痛药>亮肽素和胰蛋白酶特异性抑制剂(甲苯磺酰基-赖氨酰-氯甲基酮),表明它是一种类似于胰蛋白酶的丝氨酸蛋白酶。每10人被认为是主要的过敏原,通过皮肤测试和免疫印迹显示与80%以上的蟑螂致敏患者有IgE反应性。它可以诱导蟑螂过敏患者PBMCs培养上清液中组胺的大量释放(P <0.05)和白细胞介素4(IL-4)(P <0.05)和IL-5(P <0.05)的分泌,提示强大的致敏力。结论:从美洲疫霉分离的丝氨酸蛋白酶被证明是主要的过敏原(每10个)。它具有基于成分的过敏症诊断的潜力,将有助于阐明过敏的机制。

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